ELISA Kit for Apolipoprotein E (APOE) Oryctolagus cuniculus (Rabbit) Sandwich ELISA

Apo-E; AD2; Apoprotein; Alzheimer Disease 2(E4-Associated,Late Onset

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  • ELISA Kit for Apolipoprotein E (APOE) Packages (Simulation)
  • ELISA Kit for Apolipoprotein E (APOE) Packages (Simulation)
  • ELISA Kit for Apolipoprotein E (APOE) Results demonstration
  • SEA704Rb.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Apolipoprotein E (APOE) and the recovery rates were calculated by comparing the measured value to the expected amount of Apolipoprotein E (APOE) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 84-105 88
EDTA plasma(n=5) 90-104 101
heparin plasma(n=5) 99-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Apolipoprotein E (APOE) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Apolipoprotein E (APOE) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Apolipoprotein E (APOE) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 94-103% 86-95% 95-104% 91-105%
EDTA plasma(n=5) 84-101% 88-102% 95-102% 90-97%
heparin plasma(n=5) 98-105% 80-96% 85-98% 81-102%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Apolipoprotein E (APOE)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Apolipoprotein E (APOE). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Apolipoprotein E (APOE). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Apolipoprotein E (APOE), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Apolipoprotein E (APOE) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Cerebrospinal fluid apolipoprotein E concentration decreases after seizureSeizure: PIIS1059131109002441
  • Hepatic lipase- and endothelial lipase-deficiency in mice promotes macrophage-to-feces RCT and HDL antioxidant propertiesPubMed: 23328279
  • Expression and clinical significance of apolipoprotein E in pancreatic ductal adenocarcinomaPubmed: 23609192
  • Adipocyte Atp-Binding Cassette G1 Promotes Triglyceride Storage, Fat Mass Growth And Human ObesityPubmed:25249572
  • Frequencies of apolipoprotein E alleles in depressed patients undergoing hemodialysis – a case-control studyPubMed: 25707516
  • Culturing of HepG2 cells with human serum improve their functionality and suitability in studies of lipid metabolismPubMed: 26515253
  • Adipocyte ATP-binding cassette G1 promotes triglyceride storage, fat mass growth, and human obesityPubMed: 25249572
  • Adaptive immunity against gut microbiota enhances apoE-mediated immune regulation andreduces atherosclerosis and western-diet-related inflammation.pubmed:27383250
  • Differential effects of apoE and apoJ mimetic peptides on the action of an anti-Aβ scFv in 3xTg-AD micePubmed:30026023
  • Deletion of plasma Phospholipid Transfer Protein (PLTP) increases microglial phagocytosis and reduces cerebral amyloid-β deposition in the J20 mouse …Pubmed:29731975
  • Comparative proteomics analysis of serum proteins in gestational diabetes during early and middle stages of pregnancyPubmed: 31162828
  • Hepatic HuR modulates lipid homeostasis in response to high-fat dietPubmed: 32546794
  • Pathology and Proteomics-Based Diagnosis of Localized Light-Chain Amyloidosis in Dogs and CatsPubmed: 32880234
  • The Significance of Apolipoprotein E Measurement in the Screening of Fetal Down Syndrome33321701
  • Discovery and validation of FBLN1 and ANT3 as potential biomarkers for early detection of cervical cancer33602229
  • GP73 is a TBC-domain Rab GTPase-activating protein contributing to the pathogenesis of non-alcoholic fatty liver disease without obesity34853313
  • Iridium nanoclusters as high sensitive-tunable elemental labels for immunoassays: Determination of IgE and APOE in aqueous humor by inductively coupled …Pubmed:35378356
  • Gold nanoclusters as elemental label for the sequential quantification of apolipoprotein E and metallothionein 2A in individual human cells of the retinal …Pubmed:35361429

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